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HK1053673A1 - Methods of protein destabilization and uses thereof - Google Patents

Methods of protein destabilization and uses thereof Download PDF

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Publication number
HK1053673A1
HK1053673A1 HK03101959.4A HK03101959A HK1053673A1 HK 1053673 A1 HK1053673 A1 HK 1053673A1 HK 03101959 A HK03101959 A HK 03101959A HK 1053673 A1 HK1053673 A1 HK 1053673A1
Authority
HK
Hong Kong
Prior art keywords
linker
protein
reporter moiety
proteins
multimerized
Prior art date
Application number
HK03101959.4A
Other languages
Chinese (zh)
Inventor
H. Stack Jeffrey
Whitney Michael
B. Cubitt Andrew
A. Pollok Brian
Original Assignee
Vertex Pharmaceuticals (San Diego) Llc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Vertex Pharmaceuticals (San Diego) Llc filed Critical Vertex Pharmaceuticals (San Diego) Llc
Publication of HK1053673A1 publication Critical patent/HK1053673A1/en

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/48—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase
    • C12Q1/485—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase involving kinase
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • General Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Immunology (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Engineering & Computer Science (AREA)
  • Analytical Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

This invention is directed towards methods of destabilizing proteins in living cells, and their use for the development of novel assays. In one embodiment, the invention comprises the use of non-cleavable multimerized ubiquitin fusion proteins to destabilize a target protein, such as a reporter moiety. In one aspect of this method the constructs also comprises a linker that operatively couples the reporter moiety to the multimerized ubiquitin fusion protein. In this embodiment, enzymatic modification of the linker results in a modulation of the coupling of the reporter protein to the multimerized ubiquitin domains resulting in a change in the stability of the reporter moiety. The level of the reporter moiety in the cell can then be used as a measure of the enzymatic activity in the cell. In another embodiment the invention provides for a generalized way of coordinately regulating the cellular concentration of a plurality of target proteins. In one aspect of this method, the target proteins are operatively coupled to a ubiquitin fusion protein via a linker containing a protease cleavage site. Cleavage of the linker by a protease results in uncoupling of the target protein from the multimerized ubiquitin construct, and results in an increase in the stability and concentration of the target protein.
HK03101959.4A 2000-02-04 2001-02-02 Methods of protein destabilization and uses thereof HK1053673A1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US498098 2000-02-04
US09/498,098 US7262005B1 (en) 2000-02-04 2000-02-04 Methods of protein destabilization and uses thereof
PCT/US2001/003791 WO2001057242A2 (en) 2000-02-04 2001-02-02 Methods of protein destabilization and uses thereof

Publications (1)

Publication Number Publication Date
HK1053673A1 true HK1053673A1 (en) 2003-10-31

Family

ID=23979591

Family Applications (1)

Application Number Title Priority Date Filing Date
HK03101959.4A HK1053673A1 (en) 2000-02-04 2001-02-02 Methods of protein destabilization and uses thereof

Country Status (7)

Country Link
US (3) US7262005B1 (en)
EP (1) EP1255853A2 (en)
JP (1) JP2003534775A (en)
AU (1) AU2001234853A1 (en)
CA (1) CA2400013A1 (en)
HK (1) HK1053673A1 (en)
WO (1) WO2001057242A2 (en)

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JP4381147B2 (en) * 2002-02-25 2009-12-09 独立行政法人理化学研究所 Fluorescent protein
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AU2003218377A1 (en) * 2002-03-21 2003-10-08 California Institute Of Technology Modulation of angiogenesis through targeting of arginyl transferase (ate1)
DE602004009292T2 (en) * 2003-02-21 2009-03-19 Biotecnol S.A. USE OF CASPASES FOR THE PRODUCTION OF TIRES OF RECOMBINANT FUSION PROTEINS
CA2445420A1 (en) 2003-07-29 2005-01-29 Invitrogen Corporation Kinase and phosphatase assays
US7619059B2 (en) 2003-07-29 2009-11-17 Life Technologies Corporation Bimolecular optical probes
ATE469984T1 (en) 2003-12-01 2010-06-15 Life Technologies Corp NUCLEIC ACID MOLECULES CONTAINING RECOMBINATION SITE AND METHOD FOR USE THEREOF
US7553632B2 (en) 2004-01-22 2009-06-30 Promega Corporation Luminogenic and nonluminogenic multiplex assay
US7416854B2 (en) 2004-01-22 2008-08-26 Promega Corporation Luminogenic and nonluminogenic multiplex assay
EP1586585A1 (en) * 2004-04-14 2005-10-19 F. Hoffmann-La Roche Ag Expression system for the production of IL-15/Fc fusion proteins and their use
JP2009506772A (en) 2005-09-01 2009-02-19 プロメガ コーポレイション Cell-based luminescent and non-luminescent proteasome assays
WO2008106087A1 (en) * 2007-02-28 2008-09-04 The Brigham And Women's Hospital, Inc. Compositions and methods for identifying factors affecting protein stability
AU2008239654A1 (en) 2007-04-13 2008-10-23 Promega Corporation Luminescent live and dead cell assay
WO2013073653A1 (en) * 2011-11-18 2013-05-23 大学共同利用機関法人情報・システム研究機構 Method for inducing proteolysis in eukaryotic cell
JP2017530711A (en) * 2014-10-10 2017-10-19 ダナ−ファーバー キャンサー インスティテュート， インコーポレイテッド Methods for developing therapeutics that modify the stability of target proteins
CA2970370A1 (en) * 2014-12-24 2016-06-30 Massachusetts Institute Of Technology Crispr having or associated with destabilization domains
KR102256749B1 (en) * 2014-12-29 2021-05-25 연세대학교 산학협력단 Methods for establishing high expression cell line
CA2972990A1 (en) 2015-01-20 2016-07-28 Beatrice Tien-Yi WANG Tumor necrosis factor (tnf) superfamily receptor binding molecules and uses thereof
EP3443096B1 (en) * 2016-04-15 2023-03-01 Novartis AG Compositions and methods for selective expression of chimeric antigen receptors
US20210222164A1 (en) * 2016-06-29 2021-07-22 The Broad Institute, Inc. Crispr-cas systems having destabilization domain
US11340216B2 (en) 2016-09-13 2022-05-24 Dana-Farber Cancer Institute, Inc. Methods and compositions for the positive selection of protein destabilizers
WO2018160695A1 (en) * 2017-02-28 2018-09-07 Trustees Of Boston University Opto-genetic modulator
KR20200086278A (en) 2017-10-18 2020-07-16 노파르티스 아게 Compositions and methods for selective proteolysis

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Also Published As

Publication number Publication date
JP2003534775A (en) 2003-11-25
WO2001057242A2 (en) 2001-08-09
US20110191873A1 (en) 2011-08-04
AU2001234853A1 (en) 2001-08-14
CA2400013A1 (en) 2001-08-09
EP1255853A2 (en) 2002-11-13
WO2001057242A3 (en) 2002-06-13
US7824850B2 (en) 2010-11-02
US7262005B1 (en) 2007-08-28
US20080227129A1 (en) 2008-09-18

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